This project will study the structure and metabolism of several glycoprotein hormones related to reproduction. The isolation of the subunits of porcine FSH, and protein sequence analyses of porcine and human FSH will be carried out to resolve differences among previously published structures. Structural analyses of the separate carbohydrate side-chains of hLH, hCG and procine FSH will be done. Structure-function studies will be carried out which will help define the chemical requirements for subunit association and receptor interaction. This will include testing of deglycosylated and chemically modified subunits and synthetic fragments using bioreceptor and adenylate cyclase assays and radioimmunoassays to test completeness of association. Chemical characterization and partial sequence analysis of free ("ectopic") alpha subunit, which we are isolating from human pituitary tissue, will be done to discern the basis for the inability of the free subunit to associate in a normal manner. Region-specific antisera are being prepared using reduced, carboxymethylated subunits and their peptide fragments. These will be used as probes for detection and characterization of metabolic fragments in serum and urine. As another approach to hormone metabolism, we will introduce labelled subunits into the whole animal and identify their degradation products by Edman microsequencing methodology.